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. 2020 Aug 13;11:4061. doi: 10.1038/s41467-020-17883-1

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — a Schematic illustrating the targeting construct used in the doxycycline (dox)-inducible JUN mouse model. In this construct, rtTA is expressed at the endogenous ROSA26 promotor. With dox induction, rtTA undergoes nuclear translocation to activate the Tet-responsive element (minCMV-tet(o)) driving expression of JUN. SA splice acceptor, pA poly(A) sequence. b Representative phospho-flow-cytometry analysis for phosphorylated (phospho-) JUN (left and middle panels), and phospho-STAT5 (right panel) expression in abdominal adhesion fibroblasts isolated from JUN mice 24 h after adhesion surgery (with local induction with doxycycline at the time of adhesion formation) compared with vehicle control. n = 3 biological replicates. c Immunofluorescent assessment of phospho-JUN and phospho-STAT5 co-expression in mouse abdominal adhesion tissue. Thick white dotted line indicates edge of adhesions interface, co-expressing cells highlighted with thin white dotted lines. n = 3 biological replicates. Scale bars, 25 μm. d Quantitation of phospho-flow-cytometry analysis showing a significant decrease in phospho-JUN (top panel) and phospho-STAT5 (bottom panel) expression with application of JUN inhibitor versus vehicle control in JUN mice at 24 h. n = 3 biological replicates per condition, datapoints represent averages of technical replicates. e Representative gross images of mouse adhesions at POD 3 treated with (vehicle, DMSO) control (left panels) versus JUN inhibitor (right panels). Adhesion interface highlighted in green, structures as indicated in figures, black sutures (circled with blue dotted line) visible on inhibitor specimens are nidus for adhesion formation (these are not visible on control specimens as they are covered with bowel that is adhesed to the abdominal sidewall). n ≥ 3 biological replicates per condition per treatment. f Gross assessment (using an adhesion severity grading score established by Tsai et al. 2018¹⁹) of adhesion severity following in vivo inhibition of JUN using JUN inhibitor (T-5224) versus vehicle control in JUN (JUN expression induced with doxycycline in all JUN mice used) and wild-type mice at POD 3. n ≥ 3 biological replicates per condition per treatment. Data and error bars represent mean ± SD. *P = 0.01, **P = 0.0001, ***P = 0.004, ****P = 0.003, *****P = 0.01, ******P = 0.009, unpaired two-tailed t-test. Source data are provided as a Source Data file.